Molecular Mechanisms Underlying Endothelial Weibel-Palade Body Biogenesis and Exocytosis

内皮 Weibel-Palade 体生物发生和胞吐作用的分子机制

基本信息

批准号：
10796566
负责人：
Anish Vaibhav Sharda
金额：
$ 16.5万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-06-01 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10796566
关键词：
Adaptor Protein Complex 3 Adaptor Signaling Protein Advisory Committees Affect Albinism Binding Biogenesis Blood Blood Cells Blood Coagulation Disorders Blood Platelets Cardiovascular system Cell membrane Cells Clinical Coagulation Process Complex Cytoplasmic Granules Data Defect Dependence Disease Docking Endosomes Endothelial Cells Endothelium Environment Eukaryotic Cell Exocytosis Fellowship Five-Year Plans Genes Genetic Polymorphism Goals Growth Hematology Hemorrhage Hemostatic Agents Hemostatic function Hermanski-Pudlak Syndrome Human Immunoprecipitation Impairment Individual Israel Lysosomes Mediating Medical center Melanosomes Membrane Membrane Protein Traffic Mentors Mentorship Molecular Molecular Weight Morbidity - disease rate Neoplasm Metastasis Organelles Pathway interactions Plasma Play Postdoctoral Fellow Process Proteins Qualifying Regulation Reporting Research Research Personnel Role SNAP receptor SNAP23 gene Scientist Secondary to Secretory Vesicles Skin Pigmentation Thrombosis Training Vesicle Weibel-Palade Bodies Yeasts angiogenesis cancer cell cardiovascular risk factor career cellubrevin experimental study fluidity in vivo innovation live cell microscopy mouse model platelet function protein complex protein transport rab GTP-Binding Proteins receptor release factor syntaxin 3 trafficking trans-Golgi Network von Willebrand Disease von Willebrand Factor

项目摘要

Project Summary/Abstract Endothelium plays a vital role in hemostasis and thrombosis. It is essential to maintain blood fluidity, but it also expresses and releases numerous “factors” that regulate blood cell activation and coagulation. Among these, von Willebrand factor (vWF) is an essential plasma hemostatic factor released by the endothelium. Deficiency of mature vWF is the most common bleeding disorder in humans. On the other hand, elevated plasma levels of vWF, or abnormal concentrations of its high-molecular weight multimers, is associated with increased risk of cardiovascular morbidity. As such, understanding of the molecular mechanisms that underlie regulated release of vWF have broad implications in hemostasis and thrombosis. One factor that plays a crucial role in normal vWF exocytosis from endothelial cells is unimpaired biogenesis of its storage granules, the Weibel-Palade bodies (WPBs), which is a highly complex process that remains poorly characterized. We have previously shown that deficiency of biogenesis of lysosome-related organelle complex 2 (BLOC-2) results in impaired vWF exocytosis. Since, BLOC-2, and other related trafficking proteins, are essential for biogenesis of lysosomal-related organelles, a group of specialized granules that includes platelet dense granules and melanosomes, it seems plausible that this protein complex is also required for biogenesis of WPB. We hypothesize that BLOC-2-mediated endosomal trafficking is critical for biogenesis of WPB. We further postulate that the exocyst complex plays an essential role in WPB trafficking by 1) interacting with BLOC-2 in its role in endosomal trafficking 2) and regulating soluble NSF adaptor protein receptor (SNARE)-mediated fusion of WPBs at the plasma membrane. In Aim 1, we will characterize cargo trafficking from the endosomes to the maturing WPBs and the dependence of this pathway on BLOC-2. In Aim 2, we will evaluate the role of the exocyst complex in BLOC-2-dependent endosomal trafficking and characterize the core trafficking machinery involved in this pathway. And, finally, in Aim 3, we will evaluate the regulatory function of the exocyst complex in SNARE-mediated fusion of WPBs at the plasma membrane. The applicant, Dr. Anish Sharda, previously completed clinical and research fellowship in hematology and is currently pursuing a post-doctoral fellowship under the mentorship of Drs. Robert Flaumenhaft and Bruce Furie, who will serve as primary mentor and co-mentor, respectively. The Division of Hemostasis and Thrombosis at Beth Israel Deaconess Medical Center has a distinguished track record of scientific innovation and mentorship, and will provide an excellent environment for the applicant in pursuit of his career goals. The advisory committee of exceptional scientists that Dr. Sharda has established will bring diverse intellectual expertise to his training and scientific growth. Dr. Sharda is well-qualified to execute the proposed experiments and has presented a comprehensive five-year plan to meet his goal of becoming an independent researcher.

项目摘要/摘要内皮在止血和血栓形成中起着至关重要的作用。维持血液流动至关重要，但也表达并释放许多调节血细胞激活和凝结的“因素”。其中，冯·威尔布兰德因子（VWF）是内皮释放的必不可少的血浆止血因子。不足成熟的VWF是人类最常见的出血障碍。另一方面，血浆水平升高 vWF或其高分子重量多重浓度的异常浓度与增加的风险有关心血管发病率。因此，理解源自释放的分子机制 VWF的止血和血栓形成具有广泛的影响。在正常中起着至关重要的作用的一个因素内皮细胞中的VWF胞吐作用是其储存颗粒的无aiper的生物发生，微为微生。身体（WPB），这是一个高度复杂的过程，其特征的特征仍然很差。我们以前有表明与溶酶体相关细胞器复合物2（BLOC-2）的生物发生不足导致受损 VWF胞吐作用。由于BLOC-2和其他相关的运输蛋白对于生物发生至关重要溶酶体相关的细胞器，一组专门的颗粒，包括血小板致密颗粒和黑色素，这似乎是WPB生物发生所需的这种蛋白质复合物似乎是合理的。我们假设BLOC-2介导的内体运输对于WPB的生物发生至关重要。我们进一步假设外囊综合体在WPB运输中起着至关重要的作用1）它在内体贩运中的作用2）和调节固体NSF适配器蛋白受体（SNARE）介导的 WPB在质膜上的融合。在AIM 1中，我们将描述来自内体的货物贩运到成熟的WPB以及该途径对BLOC-2的依赖性。在AIM 2中，我们将评估依赖BLOC-2依赖性内体贩运的外囊综合体并表征了核心贩运这项路径涉及的机械。最后，在AIM 3中，我们将评估在质膜上WPB的网罗介导的融合中的外囊复合物。申请人Anish Sharda博士先前完成了血液学的临床和研究奖学金目前，正在根据DRS的精神制作的博士后奖学金。罗伯特·弗拉曼哈夫（Robert Flaumenhaft）和布鲁斯·弗里（Bruce Furie）将分别担任主要导师和同事。止血和贝丝以色列迪肯医学中心的血栓形成具有杰出的科学创新记录和Mentalship，并将为申请人追求自己的职业目标提供一个绝佳的环境。 Sharda博士确定的杰出科学家的咨询委员会将带来潜水员他的培训和科学成长的专业知识。 Sharda博士的资格良好，可以执行拟议的实验并提出了一项全面的五年计划，以实现他成为独立研究人员的目标。